PMID- 36634709 OWN - NLM STAT- MEDLINE DCOM- 20230228 LR - 20240202 IS - 1873-4995 (Electronic) IS - 0168-3659 (Print) IS - 0168-3659 (Linking) VI - 354 DP - 2023 Feb TI - Targeted GATA3 knockdown in activated T cells via pulmonary siRNA delivery as novel therapy for allergic asthma. PG - 305-315 LID - S0168-3659(23)00015-9 [pii] LID - 10.1016/j.jconrel.2023.01.014 [doi] AB - GATA3 gene silencing in activated T cells displays a promising option to early-on undermine pathological pathways in the disease formation of allergic asthma. The central transcription factor of T helper 2 (Th2) cell cytokines IL-4, IL-5, and IL-13 plays a major role in immune and inflammatory cascades underlying asthmatic processes in the airways. Pulmonary delivery of small interfering RNAs (siRNA) to induce GATA3 knockdown within disease related T cells of asthmatic lungs via RNA interference (RNAi) presents an auspicious base to realize this strategy, however, still faces some major hurdles. Main obstacles for successful siRNA delivery in general comprise stability and targeting issues, while in addition the transfection of T cells presents a particularly challenging task itself. In previous studies, we have developed and advanced an eligible siRNA delivery system composed of polyethylenimine (PEI) as polycationic carrier, transferrin (Tf) as targeting ligand and melittin (Mel) as endosomolytic agent. Resulting Tf-Mel-PEI polyplexes exhibited ideal characteristics for targeted siRNA delivery to activated T cells and achieved efficient and sequence-specific gene knockdown in vitro. In this work, the therapeutic potential of this carrier system was evaluated in an optimized cellular model displaying the activated status of asthmatic T cells. Moreover, a suitable siRNA sequence combination was found for effective gene silencing of GATA3. To confirm the translatability of our findings, Tf-Mel-PEI polyplexes were additionally tested ex vivo in activated human precision-cut lung slices (PCLS). Here, the formulation showed a safe profile as well as successful delivery to the lung epithelium with 88% GATA3 silencing in lung explants. These findings support the feasibility of Tf-Mel-PEI as siRNA delivery system for targeted gene knockdown in activated T cells as a potential novel therapy for allergic asthma. CI - Copyright (c) 2023 Elsevier B.V. All rights reserved. FAU - Kandil, Rima AU - Kandil R AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany. FAU - Baldassi, Domizia AU - Baldassi D AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany. FAU - Bohlen, Sebastian AU - Bohlen S AD - Fraunhofer Institute of Toxicology and Experimental Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) of the German Center for Lung Research (DZL), Hannover, Germany. FAU - Muller, Joschka T AU - Muller JT AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany. FAU - Jurgens, David C AU - Jurgens DC AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany. FAU - Bargmann, Tonia AU - Bargmann T AD - Fraunhofer Institute of Toxicology and Experimental Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) of the German Center for Lung Research (DZL), Hannover, Germany. FAU - Dehmel, Susann AU - Dehmel S AD - Fraunhofer Institute of Toxicology and Experimental Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) of the German Center for Lung Research (DZL), Hannover, Germany. FAU - Xie, Yuran AU - Xie Y AD - Department of Oncology, Wayne State University School of Medicine, 4100 John R St, Detroit, MI 48201, United States. FAU - Mehta, Aditi AU - Mehta A AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany; Comprehensive Pneumology Center (CPC) with the CPC-M bioArchive, Helmholtz Munich, German Center for Lung Research (DZL), Munich, Germany. FAU - Sewald, Katherina AU - Sewald K AD - Fraunhofer Institute of Toxicology and Experimental Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) of the German Center for Lung Research (DZL), Hannover, Germany. FAU - Merkel, Olivia M AU - Merkel OM AD - Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University of Munich, Butenandtstrasse 5, 81377 Munich, Germany; Comprehensive Pneumology Center (CPC) with the CPC-M bioArchive, Helmholtz Munich, German Center for Lung Research (DZL), Munich, Germany. Electronic address: Olivia.merkel@lmu.de. LA - eng GR - 637830/ERC_/European Research Council/International PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230113 PL - Netherlands TA - J Control Release JT - Journal of controlled release : official journal of the Controlled Release Society JID - 8607908 RN - 0 (RNA, Small Interfering) RN - 0 (RNA, Double-Stranded) RN - 9002-98-6 (Polyethyleneimine) RN - 0 (Transferrin) RN - 0 (GATA3 protein, human) RN - 0 (GATA3 Transcription Factor) SB - IM MH - Humans MH - RNA, Small Interfering MH - *Lung MH - *Asthma MH - RNA, Double-Stranded MH - RNA Interference MH - Polyethyleneimine MH - Transferrin MH - GATA3 Transcription Factor/genetics PMC - PMC7614985 MID - EMS185247 OTO - NOTNLM OT - Asthma OT - Cytokines OT - PCLS OT - Pulmonary delivery OT - T cell targeting OT - siRNA therapy COIS- Declaration of Competing Interest None. EDAT- 2023/01/13 06:00 MHDA- 2023/03/03 06:00 PMCR- 2024/02/01 CRDT- 2023/01/12 19:19 PHST- 2022/10/14 00:00 [received] PHST- 2022/12/20 00:00 [revised] PHST- 2023/01/06 00:00 [accepted] PHST- 2023/01/13 06:00 [pubmed] PHST- 2023/03/03 06:00 [medline] PHST- 2023/01/12 19:19 [entrez] PHST- 2024/02/01 00:00 [pmc-release] AID - S0168-3659(23)00015-9 [pii] AID - 10.1016/j.jconrel.2023.01.014 [doi] PST - ppublish SO - J Control Release. 2023 Feb;354:305-315. doi: 10.1016/j.jconrel.2023.01.014. Epub 2023 Jan 13.